貂源铜绿假单胞菌耐药性及生物学特性
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摘要
为了解貂源铜绿假单胞菌流行株基本特性,对分离的20株铜绿假单胞菌进行药物敏感性试验、血清型鉴定、ToxA基因检测、强毒株筛选及绝对致死量(LD_(100))测定。结果表明,铜绿假单胞菌分离株耐药现象比较严重,对氨苄西林、头孢呋辛、头孢曲松、头孢噻吩、头孢噻肟、头孢西丁、头孢唑林和四环素不敏感,对妥布霉素敏感率为95%;铜绿假单胞菌分离株的血清型为G型和B型,分别占85%和15%;20株铜绿假单胞菌均能检测出毒素基因ToxA;小鼠致死性试验表明,铜绿假单胞菌分离株之间毒力差异很大,少数菌株如ZHDL9、ZHDL6等均表现出较强的毒力。以菌株ZHDL9为代表株建立小鼠鼻腔感染模型,采用建立的感染模型测定得菌株ZHDL9对小鼠的LD_(100)为5×10~7 cfu。由此可见,分离的20株铜绿假单胞菌对临床常用抗生素耐药严重,对妥布霉素的敏感率为95%,提示该种抗生素可用于临床治疗水貂出血性肺炎,而菌株ZHDL9可作为生物制品研发的候选菌株。
To study the basic characteristics for epidemic strains of Pseudomonas aeruginosa isolated from mink,20 strains of Pseudomonas aeruginosa were tested for drug sensitivity,serotype identification,ToxA gene detection,screening of virulent strains and LD_(100) determination.The results showed that the isolates were resistant to ampicillin,cefuroxime,ceftriaxone,cefothiophene,cefotaxime,cefoxitin,cefazolin and tetracycline,and the susceptibility rate to tobramycin was 95%.The sera of the isolates were G-type and B-type,and the proportion was 85% and 15% respectively.ToxA gene could be detected in the isolates;the lethal test in mice showed that isolated strains appeared difference in virulence,a few strains such as ZHDL9,ZHDL6 showed a stronger virulence.Then we established the nasal infection in mice model with ZHDL9,and the absolutely lethal dose of ZHDL9 was 5×10~7 cfu.It is concluded that the 20 Pseudomonas aenginosa isolates are highly resistant to commonly used antibiotics,and the susceptibility rate to tobramycin was 95%,suggesting that this antibiotic can be used for clinical treatment for mink hemorrhagic pneumonia.Meanwhile,strain ZHDL9 can be used as a candidate strain for developing biological products.
To study the basic characteristics for epidemic strains of Pseudomonas aeruginosa isolated from mink,20 strains of Pseudomonas aeruginosa were tested for drug sensitivity,serotype identification,ToxA gene detection,screening of virulent strains and LD_(100) determination.The results showed that the isolates were resistant to ampicillin,cefuroxime,ceftriaxone,cefothiophene,cefotaxime,cefoxitin,cefazolin and tetracycline,and the susceptibility rate to tobramycin was 95%.The sera of the isolates were G-type and B-type,and the proportion was 85% and 15% respectively.ToxA gene could be detected in the isolates;the lethal test in mice showed that isolated strains appeared difference in virulence,a few strains such as ZHDL9,ZHDL6 showed a stronger virulence.Then we established the nasal infection in mice model with ZHDL9,and the absolutely lethal dose of ZHDL9 was 5×10~7 cfu.It is concluded that the 20 Pseudomonas aenginosa isolates are highly resistant to commonly used antibiotics,and the susceptibility rate to tobramycin was 95%,suggesting that this antibiotic can be used for clinical treatment for mink hemorrhagic pneumonia.Meanwhile,strain ZHDL9 can be used as a candidate strain for developing biological products.
引文
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[17] KHAJURIA A,PRAHARAJ A K,Kumar M,et al.Emergence of NDM-1 in the clinical isolates of Pseudomonas aeruginosa in India[J].Journal of Clinical & Diagnostic Research Jcdr,2013,7(7):1328-1331.
[18] MARTINO P,MARTINO J J,VILLAR J A.Hemorrhagic pneumonia caused by Pseudomonas aeruginosa in mink in Argentina[J].Revista Argentina De Microbiologia,1985,17(3):145-148.
[19] SALOMONSEN C M,THEMUDO G E,JELSBAK L,et al.Typing of Pseudomonas aeruginosa from hemorrhagic pneumonia in mink(Neovison vison)[J].Veterinary Microbiology,2013,163(1/2):103-109.
[20] CUKOR G,NOWAK N A.Affinity chromatography purification of Pseudomonas aeruginosa exotoxin A on specifically linked NAD agarose[J].Journal of Bacteriology,1982,149(3):1162-1165.
[21] SALOMONSEN C M,CHRIEL M,JENSEN T H,et al.Effect of infectious dose and season on development of hemorrhagic pneumonia in mink caused by Pseudomonas aeruginosa[J].Canadian Journal of Veterinary Research,2013,77(3):221-225.
[2] DORING G,PIER G B.Vaccines and immunotherapy against Pseudomonas aeruginosa[J].Vaccine,2008,26(8):1011-1024.
[3] ZHAO Y,GUO L,LI J,et al.Molecular epidemiology,antimicrobial susceptibility,and pulsed-field gel electrophoresis genotyping of Pseudomonas aeruginosa isolates from mink[J].Canadian Journal of Veterinary Research,2018,82(4):256-263.
[4] NIKOLAISEN N K,LASSEN D K,CHRIE M,et al.Antimicrobial resistance among pathogenic bacteria from mink (Neovison vison) in Denmark[J].Acta Veterinaria Scandinavica,2017,59(1):60.
[5] ZHANG M L,SUN C J,GU J M,et al.Salmonella Typhimurium strain expressing OprF-OprI protects mice against fatal infection by Pseudomonas aeruginosa[J].Microbiology and Immunolgy,2015,59(9):533-544.
[6] KNOX B.Pseudomonas aeruginosa som arsag til enzootiske infektioner hos mink[J].Nordisk Veterinaer Medicin,1953:731-760.
[7] JING Q,LI L,DU Y,et al.The identification,typing,and antimicrobial susceptibility of Pseudomonas aeruginosa isolated from mink with hemorrhagic pneumonia[J].Veterinary Microbiology,2014,170(3):456-461.
[8] 国家标准化管理委员会.实验动物绿脓杆菌检测方法 GB/T 14926.17—2001[S].北京:中国标准出版社,2001:63-65.
[9] 美国临床和实验室标准协会.抗菌药物敏感性试验执行标准[J].中国检验医学杂志,2012,32(3):20-179.
[10] SARLANGUE J,BRISSAUD O,LABREZE C.Clinical features of Pseudomonas aeruginosa infections[J].Archives de Pédiatrie,2006,13(S1):S13-6.
[11] WU W H,HUANG J,DUAN B Y,et al.Th17-stimulating protein vaccines confer protection against Pseudomonas aeruginosa pneumonia[J].American Journal of Respiratory and Critical Care Medicine,2012,186(5):420-427.
[12] DING B,VON SPECHT B U,LI Y Y.OprF/I-vaccinated sera inhibit binding of human interferon-gamma to Pseudomonas aeruginosa[J].Vaccine,2010,28(25):4119-4122.
[13] HARIHARAN H,COLES M,POOLE D,et al.Update on antimicrobial susceptibilities of bacterial isolates from canine and feline otitis externa[J].Canadian Veterinary Journal-revue Veterinaire Canadienne,2006,47(3):253-255.
[14] JENSEN V F,SOMMER H M,STRUVE T,et al.Factors associated with usage of antimicrobials in commercial mink(Neovison vison)production in Denmark[J].Preventive Veterinary Medicine,2016,126:170-182.
[15] 张明亮,闫新武,孙长江,等.铜绿假单胞菌F190—342-I21—83基因的克隆及其融合蛋白的原核表达[J].河南农业科学,2018,47(12):132-136,150.
[16] TAM V H,CHANG K.Prevalence,resistance mechanisms,and susceptibility of multidrug-resistant bloodstream isolates of Pseudomonas aeruginosa[J].Antimicrobial Agents and Chemotherapy,2010,54(3):1160-1164.
[17] KHAJURIA A,PRAHARAJ A K,Kumar M,et al.Emergence of NDM-1 in the clinical isolates of Pseudomonas aeruginosa in India[J].Journal of Clinical & Diagnostic Research Jcdr,2013,7(7):1328-1331.
[18] MARTINO P,MARTINO J J,VILLAR J A.Hemorrhagic pneumonia caused by Pseudomonas aeruginosa in mink in Argentina[J].Revista Argentina De Microbiologia,1985,17(3):145-148.
[19] SALOMONSEN C M,THEMUDO G E,JELSBAK L,et al.Typing of Pseudomonas aeruginosa from hemorrhagic pneumonia in mink(Neovison vison)[J].Veterinary Microbiology,2013,163(1/2):103-109.
[20] CUKOR G,NOWAK N A.Affinity chromatography purification of Pseudomonas aeruginosa exotoxin A on specifically linked NAD agarose[J].Journal of Bacteriology,1982,149(3):1162-1165.
[21] SALOMONSEN C M,CHRIEL M,JENSEN T H,et al.Effect of infectious dose and season on development of hemorrhagic pneumonia in mink caused by Pseudomonas aeruginosa[J].Canadian Journal of Veterinary Research,2013,77(3):221-225.